Mycobacteria Protocols

Volume 101 of the series Methods in Molecular Biology™ pp 31-44

Isolation of Genomic DNA from Mycobacteria

  • John T. BelisleAffiliated withDepartment of Microbiology and Immunology, University of Leicester
  • , Michael G. SonnenbergAffiliated withDepartment of Microbiology, Colorado State University

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The Mycobacterium genus is comprised of over 30 mdivldual species with a large majority being saphrophytes. However, research on these bacteria has focused primarily on the pathogens Mycobacterium bow, Mycobacterium tuberculosis, and Mycobacterium leprae, and opportunistic pathogens such as the Mycobacterium avium complex and Mycobacterium kansasil. These medically important Mycobacterium spp. have been studied extensively in terms of antigenicity (1,2), pathogenicity (3,4), and physiology (5,6), and with the development of genetic transformation techniques for mycobacteria (7), a new era of dedicated efforts to define these organisms at the genomic level was established Over the past decade, such efforts have been underscored by the isolation of mycobacterial genes encoding immunodommant proteins (2,810), virulence factors (1113) and the biosynthesis of secondary products (14,15); as well as the establishment of genome sequencing projects for M leprae and M. tuberculosis (16,17). Essential to these accomplishments was the development of efficient means to isolate high quality mycobacterial genomic DNA.